CN1094345C

CN1094345C - High dose liposomal aerosol formulations

Info

Publication number: CN1094345C
Application number: CN97196141A
Authority: CN
Inventors: J·C·沃尔德莱普; V·奈特; M·B·布莱克
Original assignee: RESEARCH DEVELOPMENT FOUDATION
Current assignee: RESEARCH DEVELOPMENT FOUDATION; Research Development Foundation
Priority date: 1996-07-03
Filing date: 1997-07-02
Publication date: 2002-11-20
Anticipated expiration: 2017-07-02
Also published as: CA2259185C; DE69729391T2; WO1998000111A1; NZ333497A; TW497974B; DE69729391D1; DE69736083T2; AU3650897A; JP2001524930A; ATE328583T1; CN1224347A; AU732124B2; ATE268163T1; EP0959874A4; EP0959874A1; DE69736083D1; CA2259185A1; EP0959874B1

Abstract

The present invention provides a high dose pharmaceutical liposome aerosol composition comprising about 12-30 mg/ml of a pharmaceutical compound, and about 130-375 mg/ml of a phospholipid starting reservoir concentration. Specifically, the present invention is drawn to anti-inflammatory glucocorticoids, immunosuppressive compounds, anti-fungal compounds, antibiotic compounds, anti-viral compounds, and anti-cancer compounds delivered via a high dose liposome aerosol composition in a phospholipid. More specifically, the invention provides a high dose cyclosporin A liposome aerosol composition comprising up to about 30 mg/ml cyclosporin A in up to about 225 mg of a phospholipid/ml starting reservoir concentration. Also provided is a high dose budesonide-liposome aerosol composition comprising up to about 15 mg/ml budesonide in up to about 225 mg of a phospholipid/ml starting reservoir concentration.

Description

High dose liposomal aerosol formulations

The background of invention

The present invention relates generally to biochemical pharmacology and pharmaceutical chemistry.Relate more specifically to the various medicines high dose liposomal aerosol formulations of (comprising cyclosporin A and budesonide).

Description of related art

For pulmonary, a lot of diseases all can utilize the aerosol delivery system to carry out the treatment of success, and this system is used for medicine is directly accumulated in the lung surface.About the administration of this mode, a large amount of instruments (for example, metered dose inhaler and Diskus) have been developed.The injecting type aerosol apparatus is used for the aerosol administration of water soluble drug and ultra micro suspension clinically; But, in the use of water-insoluble, hydrophobic compound, be restricted.

The exploitation of passing the Liposomal formulation that medicine adapts with aerosol has made the injecting type aerosol apparatus can be applied to other medicine.Utilize liposome to pass medicine and have many good qualities, comprise and water-compatible as aerosol; Can keep the Drug therapy level at pulmonary's slow release; And liposome promotes that (particularly to pulmonary alveolar macrophage) passs medicine in the cell.

Effect by aerosol location, topical therapeutic is decided by the medication amount that disease location discharged in lung; The amount that has some different key parameters decisions to discharge, thereby the treatment effect of decision aerosol preparations.The existence of for example, aerosol apparatus design and change, operating condition (as flow velocity) and auxiliary facilities (as pipe, adapter, interface, face shield etc.) is important parameter.Therefore, can improve the aerosol delivery efficiency by the proper handling of suitable aerosol apparatus instrument.The inappropriate operation of instrument and/or not good parameter can influence inhalation dose, pass the medicine position and influence therapeutic outcome.

Pharmaceutical preparation also is the key factor of regulating the aerodynamics character of aerosol delivery efficiency and drug-to-lipid body.Have now found that, the liposome that can utilize the low temperature phase transfer to prepare improves drug-to-lipid body delivery efficiency and (sees Waldrep et al., J.Of Aerosol Med.7:1994 (1994) and Waldrep et al., Int ' l J.OfPharmaceutics 97:205-12 (1993)).Other method that improves aerosol drug-to-lipid body delivery efficiency is to improve the concentration of medicine and phospholipid reservoir.Some drug-to-lipid body preparations cause the obstruction of rose with the concentration spraying that is higher than 50mg/ml; But the emptying Liposomal formulation is also arranged to (the seeing Thomas, et al., Chesy 99:168-70 (1991)) that also can successfully spray up to 150mg/ml.In addition, (output and granule (are subjected to the some effects of physicochemical property (as viscosity and surface tension) to the aerosol performance.Such variable has influenced and has passed the maximum drug-to-lipid bulk concentration that medicine adapts through the aerosol of injecting type aerosol apparatus.

Four during the last ten years, and the antiinflammatory glucocorticoid is used for treating asthma and other serious struvite pulmonary disease.Recently, the aerosol glucocorticoid treatment is used as an approach of administration more and more.At present, several different (though structural similarity) Topically active glucocorticoids are arranged, as beclometasone, budesonide, flunisolide, triamcinolone acetonide and dexamethasone, they are contained in the aerosol treatment that can be used for asthma and other struvite pulmonary disease in metered dose inhaler or the Diskus.Though uncommon, local side effects such as candidiasis and dysphonia are more common for whole body complication (forming and growth inhibited as hypothalamic pituitary axis inhibition, cataract) during with suction glucocorticoid treatment asthma, must use and assist spacer assembly.At present, do not have as yet in the U.S. and to get permission the Donisolone that is used to spray, though the ultra micro suspension of beclometasone and budesonide is used in Europe and Canada.

The present invention describes and can produce on maximum aerosol output, the granular size scope concentrated high dose cyclosporin A and budesonide-liposomal aerosol formulations in the suitableeest scope 1-3 μ m of mass median gas-kinetic diameter (MMAD).Prior art is short of on such liposomal aerosol formulations.The present invention has satisfied secular needs in this area and expectation.

The general introduction of invention

The purpose of this invention is to provide the high dose medicament-liposomal aerosol compositions that comprises about 12-30mg/ml pharmaceutical composition and the initial reservoir concentration of about 130-375mg phospholipid/ml.

In one embodiment of the invention, high dose medicament chemical compound-liposomal aerosol the compositions that comprises about 12-30mg/ml medical compounds in the≤initial reservoir concentration of about 130-375mg phospholipid/ml is provided, and wherein medical compounds is selected from antiinflammatory glucocorticoid, immunosuppressive compounds, antifungal compound, Antibiotique composition, antiviral compound and anticancer compound.

In one embodiment of the invention, provide in≤initial reservoir the concentration of about 225mg phospholipid/ml and comprised≤high dose cyclosporin A (CsA) the liposomal aerosol compositions of about 30mg/ml cyclosporin A.

In another embodiment of the present invention, provide in≤initial reservoir the concentration of about 225mg phospholipid/ml and comprised≤the high dose budesonide-liposomal aerosol compositions of about 15mg/ml budesonide (Bud).

In a preferred embodiment of the present invention, provide in≤initial reservoir the concentration of about 150mg two Laurel phosphatidyl choline (DLPC)/ml to comprise≤the high dose cyclosporin A liposomal aerosol compositions of about 20mg/ml cyclosporin A.

In a most preferred embodiment of the present invention, provide in≤initial reservoir the concentration of about 160mg two Laurel phosphatidyl choline (DLPC)/ml to comprise≤the high dose cyclosporin A liposomal aerosol compositions of about 21.3mg/ml cyclosporin A.Available other phosphatide cpd substitutes DLPC in high dose CSA Liposomal formulation.

Another aspect of the present invention is provided at≤comprise in the initial reservoir concentration of about 225mg two Laurel phosphatidyl choline (DLPC)/ml≤the high dose budesonide-liposomal aerosol compositions of about 15mg/ml budesonide.

In another preferred embodiment of the present invention, be provided at≤comprise in the initial reservoir concentration of about 200mg two Laurel phosphatidyl choline (DLPC)/ml≤the high dose budesonide-liposomal aerosol compositions of about 12.5mg/ml budesonide.Available other phosphatide cpd substitutes DLPC in high dose budesonide-Liposomal formulation.

Thereby, the invention provides high dose antiinflammatory glucocorticoid, immunosuppressive compounds, antifungal compound, Antibiotique composition, antiviral compound and the anticancer compound-liposomal aerosol compositions that in≤initial reservoir concentration of about 130-375mg phospholipid/ml, comprises about 12-30mg/ml medical compounds.

From following for the description that discloses the given preferred embodiment of the present invention of purpose, will be appreciated that of the present invention other and further content, feature and advantage.

The simple declaration of accompanying drawing

Therefore, about the acquisition of above-mentioned feature of the present invention, advantage and purpose with can understand in more detail,, can have the present invention of top simplified summary more particularly and describe by illustrated some embodiment with reference to the accompanying drawings.These accompanying drawings constitute the part of description.But what will mention is that accompanying drawing is set forth preferred embodiment of the present invention, but should not think the restriction to its scope.

Fig. 1 represents with the aerosol scattergram of Aerotech II aerosol apparatus with the flow velocity spraying high and low dose cyclosporin A-DLPC Liposomal formulation of 10 liters/minute (measured as the Andersen cascade impactor).Data (mean+SD) expressions has the classification percent (n=3 analysis) of the cyclosporin A total amount that each stage of impaction sampler of size cutout (size cut-off) (μ m) reclaims.Calculated mass intermediate value gas-kinetic diameter (MMAD) and geometric standard deviation (GSD) on logarithm-probability graph.

Fig. 2 is illustrated on people's lung analogue model with Aerotech II aerosol apparatus with 10 liters of/minute flow velocitys spraying high and low dose cyclosporin A that cyclosporin A-the DLPC Liposomal formulation is sucked.Numeric representation is the cyclosporin A that filter that 15 times Harvard respirator is connected is collected from aerosol sample at different spray times by being adjusted to tidal volume (TV) for 500ml, per minute respiration rate (BPM).

Fig. 3 represents with the aerosol scattergram of Aerotech II aerosol apparatus with the flow velocity spraying high and low dose budesonide-DLPC Liposomal formulation of 10 liters/minute (measured as the Andersen cascade impactor).Data (mean+SD) expressions has the classification percent (n=3 analysis) of the budesonide total amount that each stage of impaction sampler of size cutout (μ m) reclaims.Calculated mass intermediate value gas-kinetic diameter (MMAD) and geometric standard deviation (GSD) on logarithm-probability graph.

It is that 500ml, per minute frequency of respiration (BPM) are 15 times, record on people's lung analogue model with Aerotech II aerosol apparatus with 10 liters of/minute flow velocity spraying high and low dose budesonides that budesonide-the DLPC Liposomal formulation is sucked that Fig. 4 represents with tidal volume (TV).Numeric representation is the budesonide that filter that 15 times Harvard respirator is connected is collected from aerosol sample at different spray times by being adjusted to tidal volume (TV) for 500ml, per minute respiration rate (BPM).

Fig. 5 represents from the mimic CsA concentration of the liposome of spraying and the suction of cremophor preparation the time to be mapped.Institute is CsA-Cremophor (50mg/ml as curve; Circle), CsA-DLPC (5mg/ml; Deceive triangle) and CsA-DLPC (20mg/ml; Rhombus).

Fig. 6 represents through the lung CsA concentration behind the high dose CsA-DLPC (20mg/ml) of a respiratory time ICR mice (35g) suction spraying.

Fig. 7 represents the anti-inflammatory effect of replying that high dose Bud-DLPC attacks LPS (endotoxin) lung bronchioles eluate (BAL) leukocyte.

Fig. 8 represents diafiltration (percoll) gradient analysis of Bud-DLPC liposome.

Fig. 9 represents to increase with concentration the aerosol DLPC output (mg/ minute) of spray empty DLPC, CsA-DLPC and Bud-DLPC Liposomal formulation.(initial volume is 5ml to aerosol with the Aerotech II aerosol apparatus that is tried the water and calibrate; 10 liters of/minute flow velocitys) produce, the 4-5 of spraying minute and 6-7 minute are collected in paired samples in the AGI-4 impinger.With HPLC assay determination DLPC concentration.Institute provide data represented each indicate concentration and be subjected to test preparation, and liposome DLPC initial content (mg/ml) is mapped.

Figure 10 represents to increase spray empty DLPC, CsA-DLPC and the output (g/ minute) of Bud-DLPC Liposomal formulation with concentration.(initial volume is 5ml to aerosol with the Aerotech II aerosol apparatus that is tried the water and calibrate; 10 liters of/minute flow velocitys) produce, spray and use the reduction of analytical balance quality measurement after 10 minutes.Institute provide data represented each indicate concentration and be subjected to test preparation, and liposome DLPC initial content (mg/ml) is mapped.

Figure 11 represents to increase with concentration the aerosol CsA and the Bud output (mg/ minute) of the CsA-DLPC and the Bud-DLPC Liposomal formulation of spraying.(initial volume is 5ml to aerosol with the Aerotech II aerosol apparatus that is tried the water and calibrate; 10 liters of/minute flow velocitys) produce, the 4-5 of spraying minute and 6-7 minute are collected in paired samples in the AGI-4 impinger.Measure drug level and analyze DLPC content (Fig. 1) from aliquot sample with the HPLC analytic process.Institute provide data represented each indicate concentration and be subjected to test preparation, and liposome medicament initial content (mg/ml) is mapped.

Figure 12 represents to increase DLPC, the CsA-DLPC of emptying and viscosity (centipoise) analysis (the initial volume 10ml of Bud-DLPC Liposomal formulation with concentration; Room temperature).The data that provide are that each indicates the meansigma methods that concentration is subjected to 10 observed values of test preparation, and liposome DLPC initial content (mg/ml) is mapped.

Figure 13 represents to analyze (initial volume 7ml with DLPC, the CsA-DLPC of concentration increase emptying and surface tension (the dyne)/cm of Bud-DLPC Liposomal formulation; Room temperature).The data that provide are that each indicates the meansigma methods that concentration is subjected to 10 observed values of test preparation, and liposome DLPC initial content (mg/ml) is mapped.Sample is analysis of viscosity simultaneously.

The detailed description of invention

One of purpose of the present invention is to improve the drug effect of passing of high dose medicament compound-liposomal aerosol composition Energy. What for example, the present invention was used for improving cyclosporin A-liposomal aerosol passs the drug effect energy. At series of experiments In, determine to utilize the liposome of low phase transition temperature preparation, (contain 12 carbon atoms, saturated fat such as DLPC Fat acid side chain), can improve the output of aerosol medicine. Determine that also some sprayer is in 1-3 μ m mass median Increase the output of aerosol drug-to-lipid body in the desired magnitude range of gas-kinetic diameter (MMAD). Cyclosporin A concentration used in these early stage researchs contains 1.0mg for every milliliter of starting soln in storage liquid storehouse, DLPC is 7.5mg.

1993, people recognize need to increase cyclosporin A-liposome gas by enlarging in proportion preparation Colloidal sol output. This can finish in several ways, for example, selects more effective sprayer. By adopting Aerotech II. (ATII) sprayer (available from CIS-USA, Bedford, Mass) has enlarged the defeated of cyclosporin A in proportion Go out. This ATII increases the output of 50% aerosol approximately than in the past used Puritan Bennett Iboosj.

Second method that increases the output of aerosol drug-to-lipid body is to increase sprayer storage liquid storehouse Chinese traditional medicine and phosphorus Stock's concentration of fat. Cyclosporin A-DLPC liposome concentration successfully is increased to every ml 5mg cyclosporin A/37.5mg DLPC successfully reaches needed in 1-3 μ m mass median aerodynamic force mathematic(al) parameter (MMAD) The aerosol output of scope. Employment lung sedimentation model, this aerocolloidal the analysis showed that was A Single Intake by Inhalation 15 minutes Behind the clock, about 3.2mg cyclosporin A can be deposited in the lung in theory. The allogeneic lung transplantation of University of Pittsburgh The patient organizes research, with aerosolized cyclosporin A (being dissolved in alcohol or propane diols) patient is treated, and proves to work as The 20mg cyclosporin A reaches clinical improvements (graft rejection reverse) when passing in the lung. Use commercially available ring spore Rhzomorph A-DLPC liposome system, discharging this amount needed Inhaled Aerosol 2 hours approximately. So long every day Suction is the part trouble to the patient, and sprayer need to be store the liquid storehouse and fill 8 times again. Therefore, cyclosporin A-DLPC stock concentration must improve. But, can not spray＞the 50mg/ml liposome, this is prior art In known because bigger concentration causes the obstruction of rose.

The present invention successfully reaches the initial storehouse of 20-30mg cyclosporin A/ml:150-225mg DLPC/ml Deposit concentration. This change slightly increases granular size, and aerocolloidal MMAD rises to 2.0 μ m from 1.6 μ m, As proving with unchanged on GSD (Fig. 1) of cyclosporin A-DLPC (5mg/37.5mg). " height Dosage " output of the aerosol of 20-30mg cyclosporin A-DLPC be significantly higher than the 5mg cyclosporin A-DLPC.

Prove on simulation people lung model such as Fig. 2, sprayed 15 fens with high dose cyclosporin A-DLPC Clock disengages that the needed time of the therapeutic dose of inferring is about 45 minutes or more to the allogeneic lung transplantation patients Few. Certainly, be to determine with other cyclosporin A aerosol administrations result by other researchers during this period of time. Cyclosporin A-liposome more has when the dosage that is lower than cyclosporin A-ethanol or propane diols and toxicity in theory Effect, therefore, but respiratory time much less also as if. Cyclosporin A-DLPC is increased to about 30mg ring spore bacterium Element-225mg DLPC proves invalid.

The present invention has proved the interior high agent of 150-200mg DLPC scope of every milliliter of 20-25mg cyclosporin A The serviceability of amount cyclosporin A-DLPC liposomal aerosol is although also can consist of high agent up to 30mg/ml Amount. Available other phosphatide cpds substitute DLPC in high dose cyclosporin A-Liposomal formulation. Suitable The representative example of phosphatide cpd comprises egg PC, HSPC, two nutmegs Acyl phospholipids phatidylcholine, diolyeolyl-dipalmitoyleolyl phosphatidyl and two palmityl phosphatidyl courages Alkali.

High dose cyclosporin A-liposomal aerosol at a lot of immune-mediated PUD Ds (such as heteroplastic transplantation of the same race Thing rejection, bronchiole obturation, allergy, allergy and asthma) upward proof is useful, and at paediatrics, one-tenth Year uses different nebulizer systems proofs useful with the gerontal patient is upper. Treatment various diseases entity needs different sprays The mist time.

Cyclosporin A-the DLPC liposome is useful to the preparation high dose, because high dose cyclosporin A gas is molten Glue intends being used for the outbreak of lung transplantation thing rejection. In these researchs, patient's usefulness spraying cyclosporin A-Cremophor (50mg cyclosporin A/ml) treat. As shown in Figure 5, cyclosporin A-liposome The aerosol output of (5mg/ml and 20mg/ml) is significantly higher. Cyclosporin-the Acremophor excitant very Greatly, but this aerosol provides some clinical benefit. Therefore, cyclosporin A-liposome is in case similar trouble The person tests with it, benefit even can be more. As described about oral cyclosporin A-, cyclosporin It is effective that the A-DLPC liposome is also treated asthma.

Research of the present invention determines that also the glucocorticoid budesonide produces stable liposome, can effectively spray Aerosol in mist and the generation 1-3 μ m MMAD scope. In this stock's concentration, disengage with the ATII sprayer and to control Treat the required typical respiratory time of the every daily dose of asthma and be about 15 minutes. This be clinically accomplish easily and Practicable.

Boehringer-Ingelheim has tested the glucocorticoid liposome with a kind of spray apparatus, their utensil The every driving 20 μ l of designing requirement discharge 100-200 μ g glucocorticoid. Simple mathematical conversion shows this utensil Storage liquid storehouse in need 5,000-10,000 μ g/ml. In those experiments with this utensil, tested at ethanol and composed Budesonide in the shape agent.

Based on previous experience, in order to reach essential concentration, dense heavy-gravity suspension will be produced with Liposomal formulation.In the experiment of former use budesonide, adopt 1: 25 (budesonide: weight ratio DPLC).According to required DLPC high-load, tested various budesonides-DLPC ratio, draw with 1: 15 ratio and be advisable.Enlarging preparation then, at first is every milliliter of 5mg Bu Naide: 75mgDLPC, arrives every milliliter of 10mg budesonide at last: 150mg DLPC.Because the preparation instability expands difficulty of other glucocorticoids (beclomethasone or flunisolide) to.10mg budesonide-15mg DLPC preparation stabilization, available ATII aerosol apparatus is sprayed effectively.

Fig. 3 shows that with high dose budesonide-DLPC, the concentration increase causes aerosol particle to increase, and MMAD is increased to 2.0 μ m from 1.2 μ m.Fig. 4 shows, this high dose budesonide formulation single sucked after 15 minutes, sucks 6mg approximately or is higher than the budesonide of clinical every day of 6 times of maximal doses.Relation between the aerosol output of low dosage and high dose budesonide-DLPC is disproportionate.

Representational " high dose " budesonide-DLPC liposomal aerosol is near every milliliter of about 12.5mg budesonide, 225mg DLPC content.Available other phospholipid substitute DLPC in high dose budesonide-Liposomal formulation of the present invention.This high dose budesonide-liposomal aerosol formulations can be used for treating some struvite pulmonary disease clinically, as asthma and interstitial fibers degeneration, and the reaction of immune-mediated lung allograft rejection, bronchioles obturation, allergy and allergy.With different nebulizer systems department of pediatrics, grow up and the gerontal patient on its serviceability of proof.

The following examples purpose is to set forth various embodiments of the present invention, and is not meant to limit the present invention in any manner.

Embodiment 1

Liposomal formulation: the preparation of high dose medicament/liposome

Among the present invention for the suitableeest formulation development of various drug/lipid bodies lyophilization.The suitableeest weight ratio of having known cyclosporin A and DLPC is 1: 75.In order to determine to be suitable for the Cmax of nebulization, the aerosol that has prepared 10-30mg cyclosporin A and 75-225mg DLPC, CsA and the suitableeest weight ratio of DLPC and be 1: 75 is passed the medicine preparation.With aerosol output with suck granular size and judge and contain the 21.3mg cyclosporin A: the preparation of 160mg DLPC is the suitableeest preparation.In order to make high dose cyclosporin A-liposome reach optimization, with 100mg cyclosporin A (Sandoz Pharmaceuticals or Chemwerth Chemical Company product) and the synthetic α-lecithin 1 of 750mg, 2-two lauroyl-sn-glyceryl-3-phosphocholine (Avanti PolarLipids produces DLPC) mix.As Waldrep et al., Intl ' J.of Pharmaceutics 97:205-12 (1993) is described, in the following 37 ℃ of operations of room temperature, medicine/DLPC is mixed in the 20ml tert-butyl alcohol.After the mixing, the drug/lipid mixture moved be drawn in the vial, quick freezing, lyophilized overnight is removed the tert-butyl alcohol then, stays pulverulent mixture.Be added in the 10ml ultra-pure water of phase transition temperature (Tc) more than 25 ℃ and make multilamellar liposome, obtaining the ultimate criterion drug level is every milliliter of 1-30mg cyclosporin A: 75-225mgDLPC.Mixture was cultivated under stirring at room 30 minutes, obtained multilamellar folliculus shape liposome.Perhaps, available rotary evaporation method prepares this Liposomal formulation.Get aliquot sample and measure drug level with HPLC.Select this simple method for preparing lipidosome to be because it can easily expand as preparation in enormous quantities.

Before spraying and after the spraying, with the quality after the microscope ocular inspection Liposomal formulation swelling, promptly whether the existence of its size and drug crystallization.As O ' Riordan, et al., J.of Aerosol Med., (1996) are described in the publication, measure drug-to-lipid association (encapsulated efficient) with the percoll gradient analysis.Before spraying, the granular size of multilamellar folliculus shape high dose cyclosporin A-DLPC drug-to-lipid body preparation need not to reduce, because in spraying when (with frequent refluence), push through the shearing force that the aerosol apparatus spray-hole produced the granular size (uneven mixture is 2.2-11.6 μ m after the swelling) of medicine-liposome is further reduced.

The magnitude range of these liposomees is 271-555nm in the aerosol drops.Aqueous granule in the aerosol contains one to several liposomees.The diameter of liposome (is seen Waldrep et al., Int ' l J.of Pharmaceutics 97:205-12 (1993) less than the aqueous aerosol granule that is loaded with these liposomees.Be used for spraying after the preparation swelling in several hours.Sterilization preparation can at room temperature or in the refrigerator be preserved the several months.

For high dose budesonide-DLPC Liposomal formulation, be the optimal proportion that 1: 1 to 1: 20 different preparations are determined medicine and liposome by test budesonide-DLPC ratio.Select 1: 15 ratio (weight ratio) as the optimal proportion of high dose budesonide-DLPC preparation.This high dose formulations mixes (as top about the described method of cyclosporin A-DLPC) by 10-150mg budesonide and 150-2250mgDLPC.In the following 37 ℃ of operations of room temperature, medicine/DLPC is mixed in the 20ml tert-butyl alcohol.After the mixing, the drug/lipid mixture moved be drawn in the vial, quick freezing, lyophilized overnight is removed the tert-butyl alcohol then, stays pulverulent mixture.Add 10ml and make multilamellar liposome at the ultra-pure water of phase transition temperature (Tc) more than 25 ℃, obtaining the ultimate criterion drug level is every ml soln 1-25mg budesonide: 15-225mg DLPC.Mixture is intermittently stirred down cultivation 30 minutes in room temperature, obtain multilamellar folliculus shape liposome.Get aliquot sample and measure drug level with HPLC.Perhaps, available rotary evaporation method prepares this Liposomal formulation.

Fig. 8 represents diafiltration (percoll) gradient analysis (see O ' Riordan, et al., J.ofAerosol Med. is in the publication (1996)) of Bud-DLPC liposome.In case swelling, multilamellar folliculus budesonide-DLPC liposome at room temperature can be stablized several weeks.The sterilization goods can be stablized some months.Can add benzalkonium chloride (10mg/l) as antiseptic.

Embodiment 2

Liposomal aerosol: aerosol drug-to-lipid body treatment

In order to produce drug-to-lipid bromhidrosis colloidal sol, use Aerosol II aerosol apparatus (CIS-USA, Bedford, USA), though other commercial gas aerosol apparatus also can adopt.ATII has been specified height output, the efficient spray device (seeing Vidgren, et al., In ' l J.of Pharmaceutics 115:209-16 (1994)) that produces the suitableeest magnitude range liposomal aerosol of 1-3 μ M MMAD, is used for the administration of lung tip.To aerosol apparatus input dry air source, it is 10 liters/minute through regulating its inner dry air intake of effusion meter.The initial stock's volume of 5ml is enough to produce 15-20 minute aerosol.Need refill reservoir than the Changzhi treatment time.

Embodiment 3

Drug-to-lipid bromhidrosis sol particle size distribution

As Waldrep et al., J.of Aerosol Med.7:1994 (1994) is described, with constant granular size sampler (the Graseby Andersen Instruments Inc. on every side of Andersen 1 ACFM, Atlanta, GA), measure the aerodynamics granular size of drug-to-lipid bromhidrosis colloidal sol as people's lung simulator (Andersen).Collect the aerosol that produces from the ATII aerosol apparatus with vacuum pump (1 ACFM), each test is impacted on 8 aluminum steps (impaction on 8 aluminum stages) with 0.5 minute standard sample time.Collect on each step the aerosol drops between the 0-10 μ m after, and measure drug level (0=9.0-10.0 μ m with HPLC with 10ml ethanol or methanol-eluted fractions; 1=5.8-9.0 μ m; 2=4.7-5.8 μ m; 3=3.3-4.7 μ m; 4=2.1-3.3 μ m; 5=1.1-2.1 μ m; 6=0.65-1.1 μ m; 7=0.43-0.65m).Remove a small amount of aerosol particle with the USP artificial larynx that is connected the shock machine intake section greater than 10 μ m.Last step glass fiber collection filter.Behind the drug level of measuring with HPLC on each step, making vertical coordinate with effective cutout diameter, be mass median gas-kinetic diameter (MMAD) and the geometric standard deviation (GSD) that calculates the drug-to-lipid body on the logarithm-probability graph (Kaleida Graph 3.0) of abscissa with cumulative percentage rate (with densitometer) less than the granular size scope.Determine MMAD and GSD (seeing Waldrep et al., Int ' l J.of Pharmaceutics 97:205-12 (1993)) with being distributed in the liposome medicament content of forming in the aerocolloidal drop battle array.Drop battle array but not liposome size decision MMAD and GSD.Examine the reliability that the method is used for calculating MMAD and GSD independently with Model 3300 TSI Laser Aerosol Particle Sizer.

Embodiment 4

The estimation of inhalation dose

In order to determine the estimation inhalation dose of Bec-DLPC liposome,, collect the spraying sample on the described anthropomorphic dummy's pulmonary system of Am RevRespir Dis 143:727-37 (1991) at Smaldone et al..Use the Harvard respirator, use 15 breathings of per minute, tidal volume 500ml, aerosol sample is collected on the Whatman GF/F filter paper from ATII aerosol apparatus (10 liters/minute of flow velocitys).The basic tidal volume that records the male from the nomogram that respiratory frequency, body weight and sex are regulated is 500 (women is 450).Collect 15 minutes aerosol samples between spray phase.Extraction back is deposited on the cyclosporin A on the filter paper or the amount of budesonide with the HPLC assay determination.

Embodiment 5

The analysis of lung cyclosporin A: Solid-Phase Extraction

Carry out following step:

1. after sucking cyclosporin A-DLPC liposomal aerosol, obtain the mouse lung tissue.Add mark (1mg/ml storing solution 10 μ l) in the 10 μ gCSD.In tissue mincer or Wig-L-Bug pipe (every effective 4-5 pearl) tissue is made homogenate.

2. being organized in the 1ml ultra-pure water of homogenization extracted 1-2 minute.This volume is the amount of a tissue, if an above tissue collecting is together, and then dilution.

3. add 2ml 98% acetonitrile/2% methanol, sample is revolved mixed.

4. with sample centrifugal 20 minutes at full speed; Supernatant forwards in the clean pipe, centrifugal 10 minutes at full speed.

5. the collection supernatant in every milliliter of tissue extract, adds ultra-pure water 5ml.

6. preparation Sep-Pak C18 post (single mouse tissue Waters Sep-Pak Light) washs with 5ml 95% ethanol and 5ml ultra-pure water.Slowly add sample, with 5ml ultra-pure water and the washing of 5ml 50% acetonitrile.

7. eluent forwards in the collecting pipe, with 1ml methanol, use the 0.5ml water elution then.

8. with 1.5ml hexane wash eluent twice, discard the upper strata, thereby remove impurity.

9. temperature series being transferred under the condition of low temperature and minimum air flow is evaporated to the eluent that extracts dried with Reacti-vap.

10. rebuild in 0.3ml CSA mobile phase, sample introduction is in HPLC.

Embodiment 6

(HPLC) carries out pharmaceutical analysis with high pressure lipuid chromatography (HPLC): the budesonide analysis

HPLC measured be used for multiple purpose, measure: the budesonide content of the concentration of budesonide in the Liposomal formulation, capsule usefulness and the aerosol sample that obtains with the lung analog machine.By the concentration of HPLC assay determination budesonide, (3.9 * 150mm) posts are at room temperature measured to adopt Waters WISP 717 autopipettes and Waters Nova-Pak C18.Record the peak with variable UV/Vis wavelength detecting at the 238nm place, carry out quantitatively with WatersMillenium 2010 Chromatography Manager Version 2.15.Used mobile phase is 50: 50 ethanol/waters in these researchs, and flow velocity is (to see Andersson ﹠amp in 0.6ml/ minute; Ryrfeldt, J.PharmPharmacol 36:763-65 (1984)).Sample for analysis directly is dissolved in ethanol (lipin dissolving body).Prepare the pharmaceutical standards product from the ethanol storing solution that is stored in-80 ℃.

Embodiment 7

(HPLC) carries out pharmaceutical analysis with high pressure lipuid chromatography (HPLC): the cyclosporin A analysis

With the cyclosporin A in HPLC mensuration Liposomal formulation and the aerosol (measuring cyclosporin A content and capsule usefulness).Use Waters (Milford, MA) WISP automatic sample infusion appliance and the Supelco LC-1 post that is heated to 75 ℃ in the test.Mobile phase is 50% acetonitrile, 20% methanol and 30% water (seeing Charles et al., Ther.Drug Monitor.10:97-100 (1988)).Record the peak with variable-wavelenght detector at the 214nm place, carry out quantitatively with Waters Millenium 2010 Chromatography Manager Version 2.15.Sample for analysis directly is dissolved in methanol (lipin dissolving body).Prepare the pharmaceutical standards product from the methanol storing solution that is stored in-80 ℃.

Embodiment 8

Carrying out pharmaceutical analysis: DLPC with high pressure lipuid chromatography (HPLC) (HPLC) analyzes

The Grit ﹠amp of employing through changing; Commelin, Chem.﹠amp; The HPLC scheme of Phys.of Lipids 62:113-22 (1992).Use Waters 717 WISP automatic sample infusion appliancees and Sperisorb S5 aminoacid post (25cm * 4.6mm, 5 μ m).With acetonitrile, methanol and 10mM trifluoroacetic acid ammonium (pH4.8) (64: 28: 8 v: v: v) make mobile phase.(SEDEX 55, and Sedre France) records the peak, carry out quantitatively with WatersMillenium 2010 Chromatography Manager Version 2.15 to evaporate detector with quality.Sample for analysis directly is dissolved in ethanol or methanol (lipin dissolving body).

Embodiment 9

Drug test mouse lung model: acute inflammation test: (LPS) bronchioles lavation technology

Cause reproducible pneumonia with gram-negative cells wall lipopolysaccharide (LPS) on one's body mice.With PBsj 1600 aerosol apparatus (100 μ g/ml reservoir concentration; 60ng release dosage) the 10 fens clock times of escherichia coli 055:B5 LPS (Sigma) aerosol contact that produce cause intensive inflammatory reaction, measure this inflammatory reaction with gathering of PMN in the alveolar that responds to the chemotactic cytokine generation and (can detect in 3 hours; The peak reaction comes across and stimulated back 6 hours).Different time behind contact LPS aerosol is used methoxyflurane anesthesia, ventral aorta sacrificed by exsanguination with mice.The surgical exposure trachea carries out intubate with PE50 pipe (external diameter 0.965mm, Clay Adams).With total amount is the Hank ' s balanced salt solution (HBSS of 2.0ml; Remove Ca/Mg with EDTA) with about 1.0ml volume lung is washed 5 times.Yield typically is flushing liquor and reclaims 85%.The leukocyte of gained is counted cell centrifugation and dyeing at hematimeter.From appraisal counting, drug influence is expressed as numeration of leukocyte reduction and PMN and/or the myeloperoxidase (MPO) positive cell relevant with fixed macrophage and/or the minimizing of myeloperoxidase (MPO) negative cells number.This test is as swimming out of the quality control standards (QCS) that produces bioactive drug-to-lipid bromhidrosis colloidal sol by eliminating respiratory tract acute inflammation cell.Fig. 7 represents the anti-inflammatory effect of replying that high dose Bud-DLPC attacks LPS (endotoxin) lung bronchioles eluate (BAL) leukocyte.

Embodiment 10

Cytology: lung eluate

Pair cell prepared product on hematimeter (eluate, thymocyte cell, lymph node or splenocyte) is counted, on slide, carry out cell centrifugation (using Miles Cyto-Tek), and according to the difference of prepared product with Wright-Giemsa, May-Grunwald-Giemsa or leukocyte peroxidase stain.Under microscope oil mirror, make appraisal counting.The reduction that the biological effect of drug-to-lipid bromhidrosis colloidal sol drug-supplying system sees total white blood cells reduces with PMN or the myeloperoxidase (MPO) positive cell number purpose relevant with fixed macrophage.

Embodiment 11

The CsA that mouse lung separate tissue after the release of CsA-DLPC liposomal aerosol goes out is to antigen/short silk

Split the vitro inhibition of the lymphocyte blastogenesis that element causes

Table 1

Antigen/short silk splits plain average CPM and suppresses %

Medium 2,171

OVA 13,640

OVA+CsA?1μg/ml 2,173 99.9

Medium 517

ConA 24,341

ConA+CsA?1μg/ml 3,041 89.4

OVA=ovalbumin 250 μ g/ml; ConA=concanavalin A 1 μ g/ml;

CPM= ³3 meansigma methodss of HTdR count per minute

Liposomal aerosol is discharged into the biological activity of the CsA in the lung

In order to test, the lymphoid tissue relevant in mediastinum with bronchioles and with lung to lymph node in cause primary immune response.The Balb/c mice is put to death mice after 7 days with the local intranasal immunity (AP-OVA (80 μ g) is aided with Bordetella pertussis vaccine) of alum precipitated ovalbumin, takes out mediastinum, and isolated lymphocytes is used for analyzed in vitro.Proliferation test be included in sensitization antigen ovalbumin or with the short silk of the cell of non-specific T split element, ConA and activate with the coculture of the CsA that quantitatively goes out from the mouse lung separate tissue with solid phase extractions and HPLC after lymphocytic stimulation change.In measuring 48-72 hour ³[H}-TdR takes in the amount of DNA.With the disappearance of antigen-specific sexual stimulus or weaken or short silk splits the inhibition that inhibition that element replys proves specificity or non-specific lymphocyte activator.

Embodiment 12

Physicochemical analysis:

Surface tension and viscosity: with Tensiomat (Model 21, Fisher Scientific, Indiana PA) measures surface tension (dyne/cm).Under the condition of accurately control, the platinoiridita ring of known dimensions is treated that from survey liquid surface mentions.Will from instrument read " apparent " on duty with correction factor F, this correction factor fusion measurement ring size, fluid density and its dependent variable (by the operation instruction of manufacturer).(Gilmont Instruments, Barrington IL) measure viscosity with Gilmont Falling Ball viscometer.Viscosity (centipoise) is at room temperature measured.

Drug-to-lipid body granular size is measured: with Nicomp 370 types, Submicon Particle Sizer (Program Version 5.0, Nicomp Particle Sizing Systems, Santa Borbara, CA), to measure the granular size of drug-to-lipid liquid solution like elastic light scattering.Operation instruction according to manufacturer is measured the drug-to-lipid body sample that suspends in water, and data are expressed as the folliculus size (intensityweighted vesicle size) of density measure.As Waldrep et al., Int ' l J.of Pharmacentics 97:205-12 (1993) is described, from the outset, and spray hopper sample after 10 minutes and the aerosol sample measurement drug-to-lipid body mean diameter that reclaims with the AGI-4 impacter.

The result of Fig. 9 (mapping of DLPC concentration) shows that the aerosol output of the DLPC liposome of≤170mg/ml increases, and higher concentration is then exported minimizing.

These data expand to the CsA-DLPC liposome, obtain similar result, and 21.3mg CsA:160mg DLPC/ml obtains maximum liposomal aerosol output (Fig. 9).For the Bud-DLPC liposome, prove that its aerosol of the preparation DLPC output that contains 12.5mg Bud:187.5mg DLPC/ml is maximum.Aerosol apparatus liquid folliculus is discharged to analyze and is measured with the aerosolized quality of per minute among Figure 10 (DLPC mapping), has shown concentration dependent, the minimizing of output.

Along with the increase of liposome concentration, being accompanied by similar aerosol output increases, until critical point (Fig. 1) (mapping with drug level).Measure the aerosolized drug of CsA and Bud with the HPLC analytic process and export the Cmax (Figure 11) that has confirmed nebulization.The maximum output of CsA-DLPC liposome is at 21.3mg CsA:160mg/ml.The maximum output of Bud-DLPC is at 12.5mg Bud:187.5mg DLPC.The physico-chemical analysis proof viscosity of these Liposomal formulations (with the mapping of DLPC concentration) has parallel increase (Figure 12).The result of DLPC, Bud-DLPC and CsA-DLPC is similar.The DLPC of the ratio of viscosities sky of Bud-DLPC preparation is approximately little by 20%.It is minimum that the viscosity of CsA-DLPC keeps, and remains unchanged between 16mg CsA/120mg DLPC and 24mgCsA/180mg DLPC/ml.These results show that every kind of preparation has an aerosol with spraying to export the peak viscosity that adapts; On this threshold value, increase the drug-to-lipid bulk concentration and no longer increase output.

The result of Figure 13 (with DLPC concentration mapping) shows, CsA and Bud is added to cause in the DLPC liposome that dosage surface tension force reduces.Capillary reduction be concentration dependent and arrive one, reach the plateau value of about 100mgDLPC/ml.Do not have significantly related between the aerosol output of Liposomal formulation and the surface tension.But,, see inverse relation between surface tension and the viscosity measurement along with the increase of Liposomal formulation concentration.

Like elastic light scattering the inhomogenous initial magnitude range of the analysis showed that of drug-to-lipid body preparation is about 2.2-11.6 μ m (in Nicomp 370 upper precisions or near this upper limit) before the spraying.After the spraying, the difference that detects between preparation is for minimum.Aerosol apparatus hopper lactone plastid magnitude range is 294-502nm, with the AGI-4 impacter collect aerosol sample be 271-555nm.

High dose medicament-Liposomal formulation of selecting 10mg Bud:150mg DLPC and CsA20mg:DLPC 150mg to form is used for further aerosol research.With the analysis showed that Andersen Cascade Impactor carries out, Bud-DLPC is 2.0 μ m MMAD/1.5GSD, and CsA-DLPC is 2.0 μ m/1.8 (table 2).The analysis showed that of these preparations of on anthropomorphic dummy's lung model, being done with 15BPM and 500ml tidal volume, suction dosage 1,000 μ g Bud/ liposome every day needs 3 minutes respiratory time, and the dosage of≤5,000 μ g can suck (table 2) in 12 minutes.The result who sucks CsA-DLPC on the test lung model shows that high dose CsA-DLPC needs to suck in 4 minutes the CsA/ liposome of 5000 μ g atomizations; Suck 15,000 μ g CsA and then need 11.5 minutes (table 2).High power capacity when these results have confirmed that liposome is used for the aerosol administration.

Table 2

Spraying high dose Bud-DLPC and CsA-DLPC Liposomal formulation

Aerosol is analyzed and inhaled concentration Bud 10mg-DLPC 150mg 1,000 μ g dosage 5,000 μ g dosage 2.0 μ m MMAD ^*3 minutes 12 minutes 1.5GSD suck # and suck #CsA 20mg-DLPC 150mg 5,000 μ g dosage 15,000 μ g dosage 2.0 μ m MMAD ^*4 minutes 11.5 minutes 1.8GSD suck # and suck #

^*Andersen Cascade Impactor (meansigma methodss of 3 measurements).People's lung analogue model (15BPM/500ml TV) dosage (Bud-DLPC n=3 that # calculates with the linear regression analysis method; Analyze for CsA-DLPCn=2 time).

The present invention relates in≤initial reservoir the concentration of about 225mg phospholipid/ml, to comprise≤the high dose cyclosporin A liposomal aerosol compositions of about 30mg/ml cyclosporin A.Be more preferred from≤initial reservoir the concentration of about 160mg phospholipid/ml and comprise≤the cyclosporin A liposomal aerosol compositions of about 21.3mg/ml cyclosporin A.In general, in cyclosporin A liposomal aerosol compositions of the present invention, granular size is represented with the mass median gas-kinetic diameter, in the scope of about 1.0 μ m and about 3.0 μ m.And in cyclosporin A liposomal aerosol compositions, the ratio of cyclosporin A and phospholipid is about 1-7.5.Preferably, phospholipid is selected from egg yolk lecithin phatidylcholine, hydrogenated soya phosphatide phatidylcholine, L-Dimyristoylphosphatidylcholine, diolyeolyl-dipalmitoyleolyl phosphatidyl and dipalmitoyl phosphatidyl choline.In general, cyclosporin A liposomal aerosol compositions of the present invention is used for the treatment of immune-mediated pulmonary disease.Immune-mediated pulmonary disease like this should be selected from allograft rejection reaction, bronchioles obturation, allergy, allergy and asthma.

The invention still further relates in≤initial reservoir the concentration of about 225mg phospholipid/ml and comprise≤the high dose budesonide-liposomal aerosol compositions of about 15mg/m budesonide.High dose budesonide-liposomal aerosol compositions with in≤initial reservoir the concentration of about 225mg phospholipid/ml, comprise≤about 15mg/m budesonide is the best.Budesonide of the present invention-liposomal aerosol compositions, its granular size are represented with the mass median gas-kinetic diameter, in the scope of about 1.0 μ m and about 2.0 μ m.In budesonide-liposomal aerosol compositions, the ratio of budesonide and phospholipid is about 1-15.The representative example of phospholipid as mentioned above.Budesonide-liposomal aerosol compositions typically can be used for treating immune-mediated and struvite pulmonary disease.Immune-mediated and representative example struvite pulmonary disease so as mentioned above.Cyclosporin A is indirect inflammation-inhibiting by the blocking immunity reaction.Budesonide had not only suppressed immunoreation but also inflammation-inhibiting.These pulmonary disease can have this two kinds of compositions.

The invention still further relates to the Therapeutic Method of the individuality of immune-mediated pulmonary disease, but comprise the described individuality pharmacology of coming into operation is gone up the step of the high dose cyclosporin A liposomal aerosol compositions of acceptable dose.

The invention still further relates to and suffer from Therapeutic Method immune-mediated and individuality struvite pulmonary disease, but comprise the described individuality pharmacology of coming into operation is gone up the step of the high dose budesonide-liposomal aerosol compositions of acceptable dose.The preparation of suitable pharmaceutical compositions and administration concentration are understood easily to receiving those of ordinary skills that the present invention enlightens.

The invention still further relates in≤initial reservoir the concentration of about 225mg two Laurel phosphatidyl choline/ml and comprise≤the high dose cyclosporin A liposomal aerosol compositions of about 30mg/ml cyclosporin A.The present invention also is provided at≤comprises in the initial reservoir concentration of about 225mg two Laurel phosphatidyl choline/ml≤the high dose budesonide-liposomal aerosol compositions of about 15mg/ml budesonide.

Generally speaking, the present invention relates to comprise the high dose medicament-liposomal aerosol compositions of about 12-30mg/ml medical compounds and the initial reservoir concentration of about 130-375mg/ml phospholipid.For example, the present invention relates to the antiinflammatory glucocorticoid, immunosuppressant, antifungal compound, antibiotic, antiviral compound and the anticancer compound that disengage through the high dose liposomal aerosol compositions in phospholipid.

Any patent mentioned in this description or publication are all represented one of ordinary skill in the art's of the present invention level.To a certain extent, these patents or publication draw at this and are reference, just look like that each publication particularly, individually draws and is reference.

Those of ordinary skill in the art will readily appreciate that the present invention is very suitable for purpose, result and the advantage that realizes that the present invention mentions, and inherent those contents in institute's disclosure.The described embodiment of description and method, step, processing, molecule and particular compound are the representatives of optimum implementation, are exemplary, and do not mean that the restriction to scope of the present invention.These methods and other purposes of comprising in the spirit of the present invention that the claim scope is limited are changed to those skilled in the art and can be expected.

Claims

1. high dose medicament-liposomal aerosol compositions is characterized in that: the initial reservoir concentration that comprises 12-30mg/ml medical compounds and 130-375mg/ml phospholipid.

2. high dose cyclosporin A liposomal aerosol compositions is characterized in that: comprise≤the 30mg/ml cyclosporin A and≤the initial reservoir concentration of 225mg/ml phospholipid.

3. cyclosporin A liposomal aerosol compositions as claimed in claim 2, wherein comprise≤the 21.3mg/ml cyclosporin A and≤the initial reservoir concentration of 160mg/ml phospholipid.

4. cyclosporin A liposomal aerosol compositions as claimed in claim 2, the granular size of wherein said liposome are represented with the mass median gas-kinetic diameter, in the scope of 1.0 μ m and 3.0 μ m.

5. cyclosporin A liposomal aerosol compositions as claimed in claim 2, wherein the ratio of cyclosporin A and phospholipid is 1-7.5.

6. cyclosporin A liposomal aerosol compositions as claimed in claim 2, wherein phospholipid is selected from egg yolk lecithin phatidylcholine, hydrogenated soya phosphatide phatidylcholine, L-Dimyristoylphosphatidylcholine, dioleoyl two palmityl dioleoyl phospholipid phatidylcholine and dipalmitoyl phosphatidyl choline.

7. the described cyclosporin A liposomal aerosol of claim 2 compositions is in the purposes for preparing on the immune-mediated pulmonary disease medicine of treatment.

8. purposes as claimed in claim 7, wherein said immune-mediated pulmonary disease are selected from allograft rejection reaction, bronchioles obturation, allergy, allergy and asthma.

9. high dose budesonide-liposomal aerosol compositions, it is characterized in that: in≤initial reservoir the concentration of 225mg phospholipid/ml, comprise≤the 15mg/ml budesonide, wherein, the granular size of described liposome is represented with the mass median gas-kinetic diameter, in the scope of 1.0 μ m and 3.0 μ m.

10. budesonide as claimed in claim 9-liposomal aerosol compositions, its feature also is: comprise≤the 12.5mg/ml budesonide in the≤initial reservoir concentration of 187.5mg phospholipid/ml.

11. budesonide as claimed in claim 9-liposomal aerosol compositions, wherein the ratio of budesonide and phospholipid is 1-15.

12. budesonide as claimed in claim 9-liposomal aerosol compositions, wherein phospholipid is selected from egg yolk lecithin phatidylcholine, hydrogenated soya phosphatide phatidylcholine, L-Dimyristoylphosphatidylcholine, dioleoyl two palmityl dioleoyl phospholipid phatidylcholine and dipalmitoyl phosphatidyl choline.

13. the described budesonide of claim 9-liposomal aerosol compositions is preparing the purposes for the treatment of on immune-mediated and the inflammatory pulmonary diseases medicine.

14. the described purposes of claim 13, wherein said immune-mediated and inflammatory pulmonary diseases are selected from allograft rejection reaction, bronchioles obturation, allergy, allergy and asthma.

15. high dose cyclosporin A liposomal aerosol compositions is characterized in that: in≤initial reservoir the concentration of 160mg two Laurel phosphatidyl choline/ml, comprise≤the 21.3mg/ml cyclosporin A.

16. high dose budesonide-liposomal aerosol compositions, it is characterized in that: in≤initial reservoir the concentration of 187.5mg two Laurel phosphatidyl choline/ml, comprise≤the 12.5mg/ml budesonide, wherein, the granular size of described liposome is represented with the mass median gas-kinetic diameter, in the scope of 1.0 μ m to 3.0 μ m.

17. high dose medicament chemical compound-liposomal aerosol compositions, it is characterized in that: comprise the 12-30mg/ml medical compounds in the≤initial reservoir concentration of 130-375mg phospholipid/ml, wherein medical compounds is selected from antiinflammatory glucocorticoid, immunosuppressant, antifungal compound, antibiotic, antiviral compound and anticancer compound.